A thioredoxin‐independent fully active NADP‐malate dehydrogenase obtained by site‐directed mutagenesis
- 19 April 1993
- journal article
- Published by Wiley in FEBS Letters
- Vol. 321 (1) , 55-58
- https://doi.org/10.1016/0014-5793(93)80620-a
Abstract
A triple cysteine mutant of sorghum leaf NADP-malate dehydrogenase has been constructed by site-directed mutagenesis, combining the previously obtained mutation of the two N-terminal cysteines with the mutation of the most internal of the two C-terminal cysteines. The construct, over-expressed in E. coli, yielded an always active, dithiol-insensitive enzyme. It can be concluded that the dithiol activation of the unmodified enzyme involves a maximum of two different disulfides per subunit, and that none of the mutated cysteines is implicated in catalysis.Keywords
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